Registration Dossier

Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
19.09.2016 to 02.11.2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD 422
Version / remarks:
adopted on 28 July 2015.
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
impurity
Test material form:
solid
Specific details on test material used for the study:
98.408% 4-(Methacryloyloxy)benzophenone
0.581% Benzophenone acetate
0.054% 4-Hydroxy benzophenone

Test animals

Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone (UD), Italy.
- Age at study initiation: 8 to 9 weeks
- Weight at study initiation: 199 to 213 g for females, 168 to 197 g for males
- Fasting period before study:
- Housing:
- Diet : commercially available laboratory rodent diet (4 RF 21,Mucedola S.r.l., Via G. Galilei, 4,
20019, SettimoMilanese (MI), Italy) was offered ad libitum throughout the study
- Water (e.g. ad libitum): yes except in the case of urinalysis investigations
- Acclimation period: 19 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C±2 °C
- Humidity (%): 55%±15%
- Air changes (per hr): 15 to 20
- Photoperiod (hrs dark / hrs light): 12 hours
IN-LIFE DATES: from 28 September (day of allocation) to 15. November 2016

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
The test item was administered orally by gavage at a dose volume of 5mL/kg body weight.
Control animals received the vehicle alone at the same dose volume.
The dose was administered to each animal on the basis of the most recently recorded body weight a nd the volume administered was recorded for each animal.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis was performed to confirmthat the proposed formulation procedure was acceptable and that the stability of the formulation was satisfactory. In the present study, samples of the formulations prepared during the study were also analysed to check the concentration and homogeneity (the first a
nd the last week of treatment, where possible).
The software used for this activity was the Empower® Pro build No. 2154. Results of formulation analyses were within the acceptability limits
Details on mating procedure:
- Length of cohabitation: up to 14 days
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
- Further matings after two unsuccessful attempts: no
- After successful mating each pregnant female was caged: single
- Any other deviations from standard protocol:no
Duration of treatment / exposure:
Males
Animals were dosed once a day, 7 days a week, for 2 consecutive weeks prior to pairing,
through the mating period and thereafter through the day before necropsy (Day 49 of study).
Males were treated for a total of 48 days. Dose volumes were adjusted once per week for each
animal according to the last recorded body weight.
Females
Animals were dosed once a day, 7 days a week, for 2 consecutive weeks prior to pairing and
thereafter during pairing, post coitum and post partum periods until Day 12 post partum (for
at least 50 days). Dose volumes were adjusted once per week for each animal according to
the last recorded body weight. During the gestation period, dose volumes were calculated
according to individual body weight on Days 0, 7, 14 and 20 post coitum and Days 1, 4 and 7
post partum. Thereafter individual dose volumes remained constant.
Frequency of treatment:
Once daily
Doses / concentrationsopen allclose all
Dose / conc.:
70 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
600 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
The test item was administered orally by gavage to parental animals at 5 mL/kg body weight.
The dosages, selected in consultation with the Sponsor, were 100, 300 and 1000 mg/kg/day.

Examinations

Maternal examinations:
yes
Ovaries and uterine content:
yes
Fetal examinations:
yes
Statistics:
Standard deviationswere calculated as appropriate. For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data. The non-parametric Kruskal-Wallis analysis of variance (non-continuous variables) was used for the other parameters. Intergroup differences between the control and treated groups were assessed by the non-parametric version of the Statistical analysis of histopathological findings was carried out by means of the non-parametric Kolmogorov-Smirnov test if ‘n’ was more than 5. The criterion for statistical significance was p < 0.05. The mean values, standard deviations and statistical analysis were calculated from actual values in the computer without rounding off.

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
no effects observed
Description (incidence and severity):
Salivation was noted for a limited number of days in males and females receiving 600 mg/kg/day before pairing.
Thereafter no clinical signs were noted both in control and treated animals.
Observation of animals at removal from the cage and in an open arena (neurotoxicity assessment) did not reveal changes attributable to the test item.
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No treatment related effects were seen in animals of both sexes compared to the control group, throu
ghout the study.
Haematological findings:
no effects observed
Urinalysis findings:
no effects observed
Description (incidence and severity):
No adverse findings were observed in urinalysis assessed in males only.
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Terminal body weight was unaffected by treatment in both sexes. Some changes noted in absolute and relative organ weights in treated groups were dose unrelated and/or not accompanied by histopathological findings and therefore they were considered unrelated to treatment.
Gross pathological findings:
no effects observed
Description (incidence and severity):
There were no compound-related effects.
Neuropathological findings:
no effects observed
Description (incidence and severity):
No relevant differences were noted in all parameters investigated between control and treated groups at the end of the treatment period.
Other effects:
no effects observed
Description (incidence and severity):
Thyroid hormones in adult males
No differences in hormone levels were described between the control and the treated parental males.

Maternal developmental toxicity

Number of abortions:
no effects observed
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Description (incidence and severity):
The number of females with live pups on Day 13 post partum was: 9 in the control, 7 in the low dose (70 mg/kg/day), 9 in the mid-dose (200 mg/kg/day) and 10 in the high dose (600 mg/kg/day) groups.
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Changes in number of pregnant:
no effects observed
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Thyroid hormones in male and female pups performed on Day 13 post partum.
Some male pups of Group 3 (200 mg/kg/day) showed slight decrease of TSH (35% below mean control data). Due to the absence of dose-relation, this finding was considered unrelated to treatment.

Effect levels (maternal animals)

Key result
Dose descriptor:
NOAEL
Effect level:
600 mg/kg bw/day (actual dose received)
Remarks on result:
other: no effect observed

Maternal abnormalities

Key result
Abnormalities:
no effects observed

Results (fetuses)

Changes in sex ratio:
no effects observed
Description (incidence and severity):
Sex ratios at birth and on Day 4 post partum did not show differences between groups, when calculated as the percentage of males.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
No differences in litter data were seen between control and treated groups.
External malformations:
no effects observed
Skeletal malformations:
not specified
Visceral malformations:
not specified
Description (incidence and severity):
Cold to touch, apparently no food intake (milk), small appearance, found dead or missing pups were in general recorded as clinical signs both in control and treated pups, during the lactation period.
No relevant dose relationship was noted.

Effect levels (fetuses)

Key result
Dose descriptor:
NOAEL
Effect level:
600 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
other: no effect observed

Fetal abnormalities

Key result
Abnormalities:
no effects observed

Overall developmental toxicity

Key result
Developmental effects observed:
no

Applicant's summary and conclusion

Conclusions:
Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) for general toxicity and for reproductive and developmental toxicity was considered to be 600 mg/kg/day both for males and females.
Executive summary:

In a Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test according to OECD Guideline 422 (28 July 2015) 4-{Methacryloyloxy)benzophenone was administered to 10 Hsd: Sprague Dawley SD rats/sex/dose orally by gavage at dose levels of 0 (control), 70, 200 and 600 mg/kg bw/d. The treatment schedule included 2 weeks before pairing, during pairing, post coitum and post partum periods up to day 3 post partum. Animals were administered for approximately 5 and 8 weeks for males and females, respectively. 

 

The purpose of this study was to generate information concerning toxic effects on Sprague Dawley rats of both sexes after repeated oral dosing with 4-(Methacryloyloxy)benzophenone, as well as any effects of the test item on male and female reproductive performance, such as gonadal function, conception, parturition and lactation of offspring. The dose levels used during the study were 70, 200 and 600 mg/kg/day.

Males

One control male was sacrificed on Day 14 of the study for poor health condition. On the basis of the macroscopic and microscopic findings, the factor contributory to the death was mainly attributed to a misdosing.

Males were treated for 2 weeks prior to pairing and during pairing with females until the day before necropsy, for a total of 48 days.

During thein-lifephase, mortality check, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction tostimuli), body weight, body weight gain, food consumption and mating performance were evaluated.

Clinical pathology investigations (haematology, clinical chemistry, urinalysis), organ weights, macroscopic observations and histopathological examination were also performed. In

addition, thyroid hormone levels were determined in all adult males. The histopathological examination was performed only on control and high dose groups (5 animals/sex/group randomly selected). It included identification of the stages of the spermatogenic cycle in the

same five control and high dose males.

No relevant signs of toxicological significance were observed in treated males, apart from a transient salivation, observed for a few days in the high dose animals.

Body weights and food consumption did not show any treatment related effects. Fertility index and copulatory index were unaffected by treatment. No effects considered treatment related were observed in haematology or coagulation, clinical chemistry and urinalysis.

Hormone analysis did not showany relation to treatment. Some fluctuations in organweights, absolute and/or relative, were noted, but in the absence of histopathological correlated

findings, these changes were considered of no toxicological significance.

Females

Females were treated for 2weeks prior to pairing, during pairing and throughout the gestation and lactation periods until Day 12post partum.

During thein-lifephase, mortality check, clinical signs (including neurotoxicity assessment, motor activity and sensory reaction tostimuli), body weight, body weight gain, food

consumption and mating performance were evaluated. Clinical pathology investigations (haematology and clinical chemistry) and thyroid hormone determination in pups on Day 13 post partumwere also evaluated. The histopathological examination was carried out in five

females of control and high dose groups, selected randomly. Clinical signs of pups, as well as necropsy examination of decedent pups or pups sacrificed on Days 4 and 13 post partum (including thyroid weight) were recorded. Litter data, sex ratios and gestation length were

recorded.

No relevant signs of toxicological significance were observed. As noted for males, salivation was recorded occasionally in treated females receiving 600 mg/kg/day. Body weights and food consumption did not show changes of toxicological significance.

Concerning the reproductive parameters, no relevant differences were found in terms of mating performance including the pre-coital interval (number of days paired to sperm positive day) and the copulatory evidence (the positive identification of mating i.e. the

presence of spermand/or copulation plugin situor in the cage), as well as fertility index.

The slight changes noted at clinical chemistry investigation (haematology, clinical chemistry and coagulation) were not considered treatment related. Some fluctuations in organ weights, absolute and/or relative, were noted but in the absence of histopathological correlated.

findings, these changes were considered of no toxicological significance.

Conclusion

Based on the results of the present study, the NOAEL (No Observed Adverse Effect Level) for general toxicity and for reproductive and developmental toxicity was considered to be 600 mg/kg/day both for males and females.